title: Identification of mutations in the genome of rotavirus SA11 temperature-sensitive mutants D, H, I and J by whole genome sequences analysis and assignment of tsI to gene 7 encoding NSP3.
abstract: The complete coding sequences of the four unassigned temperature-sensitive (ts) Baylor prototype rotavirus mutants (SA11ts D, H, I and J) were sequenced by deep sequencing double-stranded RNA using RNA-seq. Non-silent mutations were assigned to a specific mutant by Sanger sequencing RT-PCR products from each mutant. Mutations that led to amino acid changes were found in all genes except for genes 1 (VP1), 10 (NSP4) and 11 (NSP5/6). Based on these sequence analyses and earlier genetic analyses, the ts mutations in gene 7, which encodes the protein NSP3, were assigned to ts mutant groups I and H, and confirmed by an in vitro RNA-binding assay with recombinant proteins. In addition, ts mutations in gene 6 were assigned to tsJ. The presence of non-conservative mutations in two genes of two mutants (genes 4 and 2 in tsD and genes 3 and 7 in tsH) underscores the necessity of sequencing the whole genome of each rotavirus ts mutant prototype.
authors.affiliation: Virologie Moléculaire et Structurale, CNRS UPR 3296, INRA USC 1358, IFR 115, Centre de Recherche de Gif, 1 Avenue de la Terrasse, 91198 Gif sur Yvette, France.
authors.forename: Patrice
authors.initials: P
authors.lastname: Vende
authors.forename: Matthieu
authors.initials: M
authors.lastname: Gratia
authors.forename: Mariela D
authors.initials: MD
authors.lastname: Duarte
authors.forename: Annie
authors.initials: A
authors.lastname: Charpilienne
authors.forename: Matthieu
authors.initials: M
authors.lastname: Saguy
authors.forename: Didier
authors.initials: D
authors.lastname: Poncet
mesh_terms: D004252:DNA Mutational Analysis; D016679:Genome, Viral; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D008969:Molecular Sequence Data; D020125:Mutation, Missense; D012367:RNA, Viral / Q000235:genetics; D012401:Rotavirus / Q000235:genetics* / Q000502:physiology / Q000528:radiation effects*; D013696:Temperature; D017361:Viral Nonstructural Proteins / Q000235:genetics* / Q000378:metabolism; D014779:Virus Replication / Q000235:genetics* / Q000528:radiation effects*
chemical_list: C082740:NSP3 protein, Rotavirus; D012367:RNA, Viral; D017361:Viral Nonstructural Proteins
keywords: Mutation; NSP3; Rotavirus; Sequencing; Temperature-sensitive